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Biomedical subjects

R Sinclair

Publications and source records attributed to R Sinclair.

At least 73 records · Page 4Linked to original sources
PubMed1992

Rapid detection of equine herpesvirus type-1 antigens in nasal swab specimens using an antigen capture enzyme-linked immunosorbent assay.

An antigen capture enzyme-linked immunosorbent assay (ELISA) was developed for the detection of equine herpesvirus type-1 (EHV-1) antigens in nasal swab specimens. The test was designed as a solid phase, amplified sandwich assay in which an EHV-1 specific monoclonal antibody was used to capture virus antigen and polyclonal antisera used to detect antigen bound to the test plates. Eight monoclonal antibodies were tested for their ability to capture virus antigen and one was selected for routine use. The sensitivity and specificity of the ELISA was compared with that of virus isolation using swabs from ponies which were experimentally infected with EHV-1. Of 72 nasal swabs collected, 32 were found to be positive by both virus isolation (VI) and ELISA, a further 15 samples were positive by VI alone, but none of the samples were positive by ELISA and negative by VI. This yielded an overall assay sensitivity of 68% and specificity of 100%. The assay proved useful for diagnosis since virus antigen was detected during the first four days post-infection which corresponded to the acute phase of disease when some clinical symptoms were apparent. In addition, the assay could be completed within one day when antibody coated plates were available.

Animals↗
PubMed1992

Influence of lidocaine on leukocyte function in the surgical wound.

The inflammatory response of the wound is mediated to a large extent by leukocytes, which play an important role in the wound healing process. Local anesthetics, which are routinely administered before minor skin surgery and for postoperative pain relief, have been shown to have diverse effects on wound healing. Local anesthetics have also been reported to induce potent inhibition of leukocytes in vitro, although their effects on leukocyte activity in the surgical wound have not been elucidated. The present study investigated the in vivo effects of lidocaine on leukocyte function in the surgical wound of rats by sampling leukocytes from hollow titanium implants. The surgical wound was treated with lidocaine or placebo after implantation of the titanium chamber and before skin closure. Leukocyte metabolic activity was measured by chemiluminescence. Cell count was analyzed in a Bürker chamber. Results showed progressive increase in leukocyte counts in the wounds of control animals and significantly lower cell counts in the wounds of lidocaine-treated animals 48 h (P less than 0.05) and 72 h (P less than 0.05) after surgery. A pronounced inhibition of the metabolic response to serum-opsonized zymosan was seen after 8 h in the lidocaine-treated animals versus controls (P less than 0.05). After 24 h, leukocyte metabolic activity decreased dramatically in the control group and remained at a low level until 72 h after surgery. In the lidocaine-treated group, the leukocyte response to zymosan remained constantly low throughout the study. The effects of lidocaine were not a result of impaired leukocyte viability.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Lignin peroxidase H2 from Phanerochaete chrysosporium: purification, characterization and stability to temperature and pH.

The wood-destroying fungus Phanerochaete chrysosporium secretes extracellular enzymes known as lignin peroxidases that are involved in the biodegradation of lignin and a number of environmental pollutants. Several lignin peroxidases are produced in liquid cultures of this fungus. However, only lignin peroxidase isozyme H8 has been extensively characterized. In agitated nutrient nitrogen-limited culture, P. chrysosporium produces two lignin peroxidases in about equal proportions. The molecular weights of these two major proteins (H2 and H8) as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were 38,500 (H2) and 42,000 (H8). The isoelectric points of these enzymes were 4.3 for H2 and 3.65 for H8. All subsequent experiments in this study were performed with H2 as it contributed the most (42%) to total activity and had the highest specific activity (57.3 U/mg). The Km values of lignin peroxidase H2 for H2O2 and veratryl alcohol were calculated to be 47 microM and 167 microM at pH 3.5, respectively. The pH optima for veratryl alcohol oxidase activity were pH 2.5 at 25 degrees C, pH 3.0 at 35 degrees C, and pH 3.5 at 45 degrees C. In the same manner the temperature optimum shifted from 25 degrees C at pH 2.5 to 45 degrees C at pH 3.5 and approximately 45-60 degrees C at pH 4.5. During storage the resting enzyme was relatively stable for 48 h up to 50 degrees C. Above this temperature the enzyme lost all activity within 6 h at 60 degrees C. At 70 degrees C all activity was lost within 10 min. The resting enzyme retained approximately 80% of its initial activity when stored at 40 degrees C for 21 h at a pH range of 4.0-6.5. Above pH 7.5 and below 4.0, the enzyme lost all activity in less than 5 h. During turnover the enzyme remained active at pH 5.5 for over 2 h whereas the enzyme activity was lost after 45 min at pH 2.5. The oxidation of veratryl alcohol was inhibited by EDTA, azide, cyanide, and by the catalase inhibitor 3-amino-1,2,4-triazole, but not by chloride. In the absence of another reducing substrate incubation of lignin peroxidase H2 with excess H2O2 resulted in partial and irreversible inactivation of the enzyme. The spectral characteristics of lignin peroxidase H2 are similar to those of other peroxidases. The suitability of lignin peroxidases for industrial applications is discussed.

Benzyl Alcohols↗
PubMed1989

The characterization of neutralizing and non-neutralizing monoclonal antibodies against equid herpesvirus type 1.

Seven monoclonal antibodies (MAbs) were produced which recognized equid herpesvirus type 1 (EHV-1). Three MAbs neutralized the subtype 1 virus (strain Army 183) in the presence of complement, but did not neutralize the subtype 2 virus (strain MD). All three MAbs immunoprecipitated an Mr 83K glycoprotein from a detergent-solubilized virion envelope preparation of the subtype 1 virus. The target antigens of the four non-neutralizing MAbs (6F11, 2A4, 1F10 and 8D9) were identified by immunoblotting against purified EHV-1 virions and had respective apparent Mr values of greater than 205K, greater than 205K, 97K and 13K.

Animals↗
PubMed1988

Topical anesthesia with lidocaine aerosol in the control of postoperative pain.

Postoperative pain was assessed in patients undergoing inguinal hernia repair. Ten patients received lidocaine aerosol in the surgical wound before skin closure, ten patients received placebo aerosol devoid of lidocaine, and ten patients were untreated. The lidocaine-treated group had significantly lower pain scores and meperidine requirements during the first postoperative day compared to the control groups. During the second day after surgery, these variables did not differ between groups. Wound anesthesia, assessed by palpation of the wound 24 h after surgery by a blinded investigator, was significantly more pronounced in the group treated with lidocaine aerosol than in the control groups. Similarly, in patients undergoing bilateral herniorraphy, wound pain following palpation was significantly reduced on the lidocaine-treated side compared to the untreated side. Patients in the group receiving lidocaine aerosol indicated less pain in connection with mobilization than untreated patients, but not compared to patients treated with placebo aerosol. Plasma substance P (SP) and beta-endorphin (BE) measured in lidocaine-treated patients and in untreated patients before and after drug administration showed no significant differences regarding SP, while BE was significantly increased 1 h after surgery in the untreated group. Plasma lidocaine concentrations were well below toxic levels. Results show that lidocaine aerosol used as topical anesthetic in the surgical wound is simple to use, and results in a long-lasting reduction of pain after a single administration. Moreover, postoperative mobilization is facilitated, and the requirement for postoperative analgesics is reduced. Wound healing was normal, and no adverse reactions to lidocaine were reported.

Adult↗
PubMed1988

Responses from area 3b of somatosensory cortex to textured surfaces during active touch in primate.

(1) The purpose of this experiment was to characterize the responses of neurons in somatosensory cortex while the hand was actively moved (stroked) across a textured surface. Surfaces consisted of horizontal gratings that varied by spatial period or ridge-groove ratio (roughness). Surfaces were attached to rectangular blocks. TOP and BOTTOM halves of each block could contain surfaces of different roughness. (2) Velocity and force of the stroke were behaviorally constrained within certain limits and continuously measured and recorded during the stroke. (3) Response samples for each neuron were obtained for repeated presentations of each surface. Statistical analyses consisted of analysis of variance and t tests across surfaces on the data of each neuron, and summary statistics on groups of neurons with similar response characteristics. The interaction effects of behavioral variables (velocity and force) were examined and found not to be significant. (4) The sample mainly consisted of rapidly adapting neurons in area 3b of somatosensory area I (SI). Three main response types were found: (a) GRADED cells showed a monotonic increase in firing rate to increasingly rougher surfaces. This effect was seen in one-third of cells studied and is consistent with other reports. These cells seem to code roughness in the magnitude of their response. (b) In some cells, response to a BOTTOM surface depended on the roughness of the preceding TOP surface. This is analogous to contrast in the visual system. These CONTRAST cells are a novel finding in the somatosensory system. (c) Some cells only responded to surfaces that were completely smooth. These "OFF"-response-type cells were seen in proximity to other cells that responded in a reciprocal fashion to surfaces with ridges, but not to smooth surfaces. SMOOTH cells did not respond to punctate or passively applied stimuli, and therefore could not be classified by adaptation of the responses. (5) An increase in firing rate as spatial period (roughness) increases (with a constant ratio of ridge to groove) seems contrary to vibratory models of texture perception. As spatial period increases, temporal frequency decreases, and thus "tuned" cells should show a decreased response rate. Yet GRADED cells showed an increased response. In addition, response varied on surfaces with different groove size, where spatial period, and thus temporal period, was constant. This suggests that in rapidly adapting neurons, at least for these simple surfaces, texture is coded by the magnitude of the firing rates rather than by its temporal fidelity.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1982

A clinical assessment of fluorescence polarisation changes in lymphocytes stimulated by phytohaemagglutinin (PHA) in malignant and benign diseases.

The modified double-zone SCM technique, developed in these laboratories as an in vitro test for cancer, is based on the differential response to PHA of lymphocytes harvested from two regions of a Ficoll-Triosil gradient. Lymphocyte responses are measured by changes in intracellular fluorescein fluorescence polarisation. We report its continued clinical evaluation in patients hospitalised with malignant and non-malignant diseases, including a blind trial of 78 patients with disorders of the gastrointestinal tract, breast and lung. Overall "false' negative and "false' positive rates from 336 blood samples were 1.8 and 3.3% respectively.

Adult↗
PubMed1981

Glycoprotein biosynthesis in animal cells grown in suspension culture. Assembly of lipid-linked saccharides and formation of protein-bound 'high-mannose' oligosaccharides.

Glycoprotein biosynthesis was studied with mouse L-cells grown in suspension culture. Glucose-deprived cells incorporated [3H]mannose into 'high-mannose' protein-bound oligosaccharides and a few relatively high-molecular-weight lipid-linked oligosaccharides. The latter were retained by DEAE-cellulose and turned over quite slowly during pulse--chase experiments. Increased heterogeneity in size of lipid-linked oligosaccharides developed during prolonged glucose deprivation. Sequential elongation of lipid-linked oligosaccharides was also observed, and conditions that prevented the assembly of the higher lipid-linked oligosaccharides also prevented the formation of the larger protein-bound 'high-mannose' oligosaccharides. In parallel experiments, [3H]mannose was incorporated into a total polyribosome fraction, suggesting that mannose residues were transferred co-translationally to nascent protein. Membrane preparations from these cells catalysed the assembly from UDP-N-acetyl-D-[6-3H]glucosamine and GDP-D-[U-14C]mannose of polyisoprenyl diphosphate derivatives whose oligosaccharide moieties were heterogeneous in size. Elongation of the N-acetyl-D-[6-3H]glucosamine-initiated glycolipids with mannose residues produced several higher lipid-linked oligosaccharides similar to those seen during glucose deprivation in vivo. Glucosylation of these mannose-containing oligosaccharides from UDP-D-[6-3H]glucose was restricted to those of a relatively high molecular weight. Protein-bound saccharides formed in vitro were mainly smaller in size than those assembled on the lipid acceptors. These results support the involvement of lipid-linked saccharides in the synthesis of asparagine-linked glycoproteins, but show both in vivo and in vitro that protein-bound 'high-mannose' oligosaccharide formation can occur independently of higher lipid-linked oligosaccharide synthesis.

Animals↗
PubMed1981

Fluoride inhibition of the hydro-osmotic response of the toad urinary bladder to antidiuretic hormone.

The hydro-osmotic response of the toad bladder to antidiuretic hormone and cyclic AMP was inhibited by the methoxyflurane metabolite, fluoride. The osmotic transfer of water in the absence of hormone was unaffected by fluoride as was the hydroosmotic response due to hypertonicity of the serosal bathing media. Osmotic water movements across N-ethylmaleimide-"fixed" vasopressin or cyclic AMP-stimulated bladders were likewise unchanged by fluoride, suggesting that fluoride is exerting an action subsequent to the endogenous formation of cyclic AMP but before the final effector mechanism. Fluoride increased intracellular cyclic AMP concentrations even in the presence of added hormone. Fluoride suppressed calmodulin activity and prevented its activation of phosphodiesterase. Fluoride had no effect on oxygen consumption of toad urinary bladder cells but reduced lactate formation and anerobic metabolism. This decrease in the glycolytic energy source did not contribute to the inhibition of the hormonal response since 2-deoxyglucose was without effect on hormonal mediated osmotic-water flow. It is postulated that the fluoride-induced polyuria after methoxyflurane anesthesia may be due in part to the ability of fluoride to interfere with calcium and calmodulin-initiated processes (other than phosphodiesterase activity) that may occur in the stimulus-reabsorption coupling response of antidiuretic hormone.

Aerobiosis↗
PubMed1980

Glucose metabolism and dehydrogenase activities in the cytosol and mitochondria of mouse LS cells in chemostat culture.

Energy metabolism has been examined in mouse LS cells growing under steady-state conditions in chemostat culture. The metabolic quotient of glucose oxidized (glucose consumed, but not fermented) remained constant, independent of growth rate between cell doubling times of 6 days and 1.2 days. Specific activities of cytochrome oxidase and malate dehydrogenase in the mitochondria remained constant at different growth rates, in accord with the constancy of the glucose oxidation rate. Cytosolic malate dehydrogenase activity was about fourfold greater than the mitochondrial isozyme. The steady-state rate of lactate production fluctuated because of technical limitations but correlated well with cytosolic lactate dehydrogenase activity.

Animals↗
PubMed1980

Phosphorylation state of mitochondria and cytosol of 3T3 mouse cells.

Preconfluent quiescent 3T3 mouse cells, stimulated into growth with 10% serum, showed little change in their content of ATP per unit protein, or in phosphorylation state (ATP/ADP ratio) which remained near 3.0 during the initial 24 hours. In quiescent, completely confluent cells, the ATP/ADP ratio was reduced to 1.3, but increased to the normal value after 20 hours stimulation with 10% serum. These changes occurred in the cell cytoplasm; in the mitochondria the ATP/ADP ratio remained near 0.5 in both inhibited and growing cells.

Adenine Nucleotides↗
PubMed1979

Toxic rash associated with high dose methotrexate therapy.

Fifteen rashes were observed in thirteen patients in association with high dose methotrexate therapy. The lack of recurrence of the rash with further treatment courses and the association of the rash with other toxic manifestations and with larger doses of methotrexate suggests a toxic mechanism. Rashes have frequently been reported in association with dose methotrexate therapy (Van Scott, Auerbach & Weinstein, 1964; Leone, Albala & Rege, 1968; Mitchell et al., 1968; Capizzi et al., 1970; Jaffe et al., 1973; Rosen, Suwansirikul & Kwon, 1974; Pratt et al., 1975; Jaffe & Traggis, 1975; Ensminger & Frei, 1977; Stoller et al., 1977). They include perifolliculitis, transient erythema progressing to maculopapular eruptions, occasionally desquamating, sloughing over pressure areas, reactions confined to radiation portals, exacerbation of acne, photosensitivity and rarely urticaria. Relatively few reports sufficiently document the incidence or types of rash. It has been suggested that the rash is allergic in nature (Mitchell et al., 1968) or a toxic phenomenon (Djerassi et al., 1972; Djerassi & Kim, 1976), possibly related to drug effects on small vessels (Van Scott, 1963).

Erythema↗